Need colorimetric assay to detect PCR products.

Daniel Kim dkim at nmsu.edu
Tue Aug 8 12:05:17 EST 1995


WRT quick assay for the presence of PCR products:

I believe one can simply add ethidium bromide to the reaction mixes, and 
assay for UV-induced fluorescence.  The ethidium bromide does not 
interfere with the PCR reaction.  Fluorescence from primers and dNTPs is 
very low, while fluorescence increases with the presence of dsDNA 
products.  It is even possible to assay the progress of the reaction in 
real time, with the right equipment.  If one did a 96-well array of 
reactions (e.g. in a PE 9600 machine), including ethidium bromide, then 
placed the array of tubes on a transilluminator, one could photograph the 
array from the top, giving a nice +/- assay of DNA synthesis in each tube.





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